Two‐factor requirement for murine megakaryocyte colony formation

Williams, Neil; Eger, Richard; Jackson, Heather; Nelson, Donald J.

doi:10.1002/jcp.1041100116

Cited by 164 publications

(63 citation statements)

References 20 publications

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“…Short-term assays for granulocyte progenitor cells (CFU-GM) and megakaryocyte progenitor cells (CFU-MK) were performed as previously described (14, 15). Briefly, bone marrow cells were cultured as previously described using conditioned medium from WEHI-3 and mouse lung conditioned medium (16,17). Before plating, the bone marrow cells were generally incubated at 370C for 30-60 min at -5 X 106 cells/ml in McCoy's SA medium containing 10% fetal calf serum (FCS).…”

Section: Methodsmentioning

confidence: 99%

“…Marrow cells were cultured at 30-100 X 104 cells/ml in 35-mm petri dishes (Becton-Dickinson & Co.) in modified McCoy's SA medium containing 10% FCS and 0.25% Bactoagar. After incubation (7 d, 370C, 7% C02, 100% humidity), petri dishes were removed from the incubator, dried, and stained in situ for acetylcholinesterase activity (18). Colonies were counted at a magnification of 40.…”

Section: Methodsmentioning

confidence: 99%

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Extracellular matrix promotes the growth and differentiation of murine hematopoietic cells in vitro.

Campbell¹,

Wicha²,

Long³

1985

J. Clin. Invest.

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We have developed a long-term culture system in which murine marrow cells are cultured on a complex extracellular matrix (ECM) that is derived from marrow and extracted with guanidine hydrochloride and dithiothreitol. Marrow cultures were established with fresh murine marrow cells and recharged at 2 wk (week 0). Phase microscopy showed a dramatically increased adherent cell layer development on ECM compared with controls within a week after recharge. By electron microscopy, this adherent layer was composed of numerous reticular cells apparently attached to the ECM which extended cytoplasmic projections to the surrounding hematopoietic cells. Adherent cellularity on ECM-coated dishes increased to 30 times the control values by week 2. Cumulative suspension cells on ECM dishes were eight times controls. ECM influenced both hematopoietic progenitor cell proliferation and differentiation. Adherent colony-forming unit-granulocyte/macrophage and colony-forming unit-megakaryocyte were >30 and 15 times the control values, respectively, by week 2 (P < 0.05). There were more mature granulocytic and megakaryocytic cells in ECM-coated dishes than in controls at all time points. This new culture system directly demonstrates that ECM is an important component of the hematopoietic microenvironment.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Extracellular matrix promotes the growth and differentiation of murine hematopoietic cells in vitro.

Campbell¹,

Wicha²,

Long³

1985

J. Clin. Invest.

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“…The effect of GM-CSF on reticulocyte numbers (17) may depend on erythropoietin, a cofactor that was suppressed in our studies by red cell transfusion. The inconsistent effect of GM-CSF on platelet numbers might also reflect variable amounts ofa second factor, e.g., thrombopoietin (22). The increase in platelet count observed after GM-CSF in the two animals with prolonged pancytopenia and bone marrow hypoplasia support an ability of this factor to augment platelet production.…”

Section: Discussionmentioning

confidence: 92%

Recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSF) shortens the period of neutropenia after autologous bone marrow transplantation in a primate model.

Nienhuis¹,

Donahue²,

Karlsson³

et al. 1987

J. Clin. Invest.

166

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The effect of granulocyte-macrophage colony-stimulating factor (GM-CSF) on hematopoietic reconstitution after autologous bone marrow transplantation was evaluated in a primate model. Animals were given a continuous intravenous infusion of recombinant human GM-CSF for several days both before and after transplantation or only after the transplant procedure. Marrow ablation was accomplished by total body irradiation. In both groups of animals, the neutrophil count reached 1,000/mm3 by 8-9 d posttransplant compared with an interval of 17 and 24 d for two concurrent controls. After withdrawal of GM-CSF, neutrophil counts fell to values comparable to those observed in untreated controls. Accelerated recovery of platelet production was also observed in four of the five animals. Two additional animals were initially given GM-CSF several weeks posttransplantation because of inadequate engraftment. Prompt and sustained increases in neutrophil and platelet counts were observed. We conclude that GM-CSF may be useful in accelerating bone marrow reconstitution.

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“…Williams et al have suggested that separate regulatory molecules influence early and late megakaryocytopoiesis. They demonstrated two regulatory factors, which had on average molecular weight of 35,000 (megakaryocytic colony stimulating factor) and 100,000 (potentiating factor) in WEHI-3 murine megakaryocytic leukemic cell conditioned medium (WEHI-3 CM), and that both activities were necessary for maximal colony development and detection (22). Serum thrombopoietin or thrombopoietic stimulating factor derived from the supernatant of a human embryonic kidney cell line has been considered as a potentiator of late megakaryocytopoiesis (23,24).…”

Section: Discussionmentioning

confidence: 99%

Effect of gangliosides on murine megakaryocytopoiesis in a liquid culture system.

Watanabe

Taniguchi

Fukamachi

et al. 1988

Cell Struct. Funct.

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ABSTRACT.Bovine brain gangliosides were applied to primary cultures of murine bone marrow cells to examine the role of gangliosides in development of megakaryocytes.Megakaryocytes in the cultures were detected by staining for a cytoplasmic enzyme, acetylcholinesterase, and divided into two types, 1)immature megakaryocytes which were stained less intensely, and 2) mature ones which were stained intensely. A medium containing total ganglioside fraction from bovine brain increased the number of both immature and mature megakaryocytes in the presence of pokeweed mitogen-stimulated murine spleen cell conditioned medium. Between the two cell types, the number of the mature cells was more significantly increased than the immature cells. The ganglioside GDla could substitute for the total ganglioside mixture. The results suggested that bovine brain gangliosides potentiated both megakaryocytic proliferation and maturation in vitro.

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Two‐factor requirement for murine megakaryocyte colony formation

Cited by 164 publications

References 20 publications

Extracellular matrix promotes the growth and differentiation of murine hematopoietic cells in vitro.

Extracellular matrix promotes the growth and differentiation of murine hematopoietic cells in vitro.

Recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSF) shortens the period of neutropenia after autologous bone marrow transplantation in a primate model.

Effect of gangliosides on murine megakaryocytopoiesis in a liquid culture system.

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